Synergistic Induction of Apoptosis by the Combination of an Axl Inhibitor and Auranofin in Human Breast Cancer Cells = Synergistic Induction of Apoptosis by the Combination of an Axl Inhibitor and Auranofin in Human Breast Cancer Cells

다국어 초록 (Multilingual Abstract)

Axl receptor tyrosine kinase has been implicated in cancer progression, invasion, and metastasis in various cancer types. Axl overexpression has been observed in many cancers, and selective inhibitors of Axl, including R428, may be promising therapeutic agents for several human cancers, such as breast, lung, and pancreatic cancers. Here, we examined the cell growth inhibition mediated by R428 and auranofin individually as well as in combination in the human breast cancer cell lines MCF-7 and MDAMB- 231 to identify new advanced combination treatments for human breast cancer. Our data showed that combination therapy with R428 and auranofin markedly inhibited cancer cell proliferation. Isobologram analyses of these cells indicated a clear synergism between R428 and auranofin with a combination index value of 0.73. The combination treatment promoted apoptosis as indicated by caspase 3 activation and poly (ADP-ribose) polymerase cleavage. Cancer cell migration was also significantly inhibited by this combination treatment. Moreover, we found that combination therapy significantly increased the expression level of Bax, a mitochondrial proapoptotic factor, but decreased that of the X-linked inhibitor of apoptosis protein. Furthermore, the suppression of cell viability and induction of Bax expression by the combination treatment were recovered by treatment with N-acetylcysteine. In conclusion, our data demonstrated that combined treatment with R428 and auranofin synergistically induced apoptosis in human breast cancer cells and may thus serve as a novel and valuable approach for cancer therapy.

참고문헌 (Reference)

1 Tang, B., "Upregulation of Akt/NF-κB-regulated inflammation and Akt/Bad-related apoptosis signaling pathway involved in hepatic carcinoma process : suppression by carnosic acid nanoparticle" 11 : 6401-6420, 2016

2 Yao, H., "Triple-negative breast cancer : is there a treatment on the horizon" 8 : 1913-1924, 2017

3 Cuadrado, A., "Transcription factors NRF2 and NF-κB are coordinated effectors of the Rho family, GTP-binding protein RAC1 during inflammation" 289 : 15244-15258, 2014

4 Cuadrado, A., "Transcription factor NRF2 as a therapeutic target for chronic diseases : a systems medicine approach" 70 : 348-383, 2018

5 Leconet, W., "Therapeutic activity of anti-Axl antibody against triple-negative breast cancer patient-derived xenografts and metastasis" 23 : 2806-2816, 2017

6 Goyette, M. A., "The receptor tyrosine kinase Axl is required at multiple steps of the metastatic cascade during HER2-positive breast cancer progression" 23 : 1476-1490, 2018

7 Koorstra, J. B., "The Axl receptor tyrosine kinase confers an adverse prognostic influence in pancreatic cancer and represents a new therapeutic target" 8 : 618-626, 2009

8 Gelmon, K., "Targeting triple-negative breast cancer : optimising therapeutic outcomes" 23 : 2223-2234, 2012

9 Vidula, N., "Targeted therapy for metastatic triple negative breast cancer: the next frontier in precision oncology" 8 : 106167-106168, 2017

10 Jin, S., "Targeted drug delivery for breast cancer treatment" 8 : 143-153, 2013

11 이주은, "Synergistic induction of apoptosis by combination treatment with mesupron and auranofin in human breast cancer cells" 대한약학회 40 (40): 746-759, 2017

12 정주희, "Role of G Protein-Coupled Estrogen Receptor in Cancer Progression" 한국독성학회 35 (35): 209-214, 2019

13 Miller, M. A., "Reduced proteolytic shedding of receptor tyrosine kinases is a post-translational mechanism of kinase inhibitor resistance" 6 : 382-399, 2016

14 Holland, S. J., "R428, a selective small molecule inhibitor of Axl kinase, blocks tumor spread and prolongs survival in models of metastatic breast cancer" 70 : 1544-1554, 2010

15 Kapur, A., "Oxidative stress via inhibition of the mitochondrial electron transport and Nrf-2-mediated anti-oxidative response regulate the cytotoxic activity of plumbagin" 8 : 1073-, 2018

16 Lu, K., "NRF2 induction supporting breast cancer cell survival is enabled by oxidative stressinduced DPP3-Keap1 interaction" 77 : 2881-2892, 2017

17 Sun, S. Y, "N-acetylcysteine, reactive oxygen species and beyond" 9 : 109-110, 2010

18 Shamas-Din, A., "Mechanisms of action of Bcl-2 family proteins" 5 : a008714-, 2013

19 Zhang, P., "Loss of Kelch-like ECH-associated protein 1 function in prostate cancer cells causes chemoresistance and radioresistance and promotes tumor growth" 9 : 336-346, 2010

20 Maes, M. E., "Live-cell imaging to measure Bax recruitment kinetics to mitochondria during apoptosis" 12 : e0184434-, 2017

21 Chun Chu, "Involvement of Estrogen Receptor-α in the Activation of Nrf2-Antioxidative Signaling Pathways by Silibinin in Pancreatic β-Cells" 한국응용약물학회 28 (28): 163-171, 2020

22 Katz, H., "Immunotherapy in triple-negative breast cancer" 35 : 13-, 2017

23 Shaw, I. C., "Gold-based therapeutic agents" 99 : 2589-2600, 1999

24 Gay, C. M., "Giving AXL the axe : targeting AXL in human malignancy" 116 : 415-423, 2017

25 Wang, C., "Gas6/Axl axis contributes to chemoresistance and metastasis in breast cancer through kat/GSK-3β/β-catenin signaling" 6 : 1205-1219, 2016

26 Tallarida, R. J., "Drug synergism : its detection and applications" 298 : 865-872, 2001

27 Cazanave, S. C., "Degradation of Keap1 activates BH3-only proteins Bim and PUMA during hepatocyte lipoapoptosis" 21 : 1303-1312, 2014

28 Bayat Mokhtari, R., "Combination therapy in combating cancer" 8 : 38022-38043, 2017

29 Probst, B. L., "Cancer cell growth is differentially affected by constitutive activation of NRF2 by Keap1 deletion and pharmacological activation of NRF2 by the synthetic triterpenoid, RTA 405" 10 : e0135257-, 2015

30 Nagini, S., "Breast cancer : current molecular therapeutic targets and new players" 17 : 152-163, 2017

31 Pawlowski, J., "Bax-induced apoptotic cell death" 97 : 529-531, 2000

32 Oommen, D., "BRCA1 deficiency increases the sensitivity of ovarian cancer cells to auranofin" 784-785 : 8-15, 2016

33 Zhang, Y. X., "Axl is a potential target for therapeutic intervention in breast cancer progression" 68 : 1905-1915, 2008

34 Varghese, E., "Auranofin, an anti-rheumatic gold compound, modulates apoptosis by elevating the intracellular calcium concentration in MCF-7 breast cancer cells" 6 : 2243-2258, 2014

35 Fiskus, W., "Auranofin induces lethal oxidative and endoplasmic reticulum stress and exerts potent preclinical activity against chronic lymphocytic leukemia" 74 : 2520-2532, 2014

36 Lee, W. P., "Akt is required for Axl-Gas6 signaling to protect cells from E1A-mediated apoptosis" 21 : 329-336, 2002

37 Katoh, H., "Activation of Rac1 by RhoG regulates cell migration" 119 : 56-65, 2006

38 Mine, N., "Activation of Nrf2 pathways correlates with resistance of NSCLC cell lines to CBP501 in vitro" 13 : 2215-2225, 2014

39 Faix, J., "A dual role model for active Rac1 in cell migration" 4 : 110-115, 2013

40 Sachan Richa, "A New Histone Deacetylase Inhibitor, MHY4381, Induces Apoptosis via Generation of Reactive Oxygen Species in Human Prostate Cancer Cells" 한국응용약물학회 28 (28): 184-194, 2020